Abstract
Initiation of transcription by RNA polymerase II from a promoter region on DNA requires the assembly of several initiation factors to form a preinitiation complex. Assembly of this complex1,2 is initiated by the binding of the transcription factor TFIID, composed of the TATA-box binding protein (TBP) and TBP-associated factors (TAFIIs), to the promoter. We have now characterized an immunopurified TFIID complex which we unexpectedly find contains the cleavage–polyadenylation specificity factor (CPSF), one of the factors required for formation of the 3′ end of messenger RNA3,4. CPSF is brought to the preinitiation complex by TFIID, but after transcription starts, CPSF dissociates from TFIID and becomes associated with the elongating polymerase. We also show that overexpression of recombinant TBP in HeLa cells decreases polyadenylation without affecting the correct initiation of transcription of the reporter gene. This indicates that, owing to incomplete assembly of TFIID on recombinant TBP, CPSF is not brought to the promoter and therefore polyadenylation becomes less efficient. Our observations have thus revealed a link between transcription initiation and elongation by RNA polymerase II and processing of the 3′ end of mRNA.
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Acknowledgements
We thank P. Chambon for his continuous support; A. Staub for peptide sequencing; J. M. Chipoulet for DE0.35 fractions; D. Boam, A. Bertolotti, I. Davidson, V. Dubrovskaya, X. Jacq, A.C.Lavigne, G. Mengus and M. Vigneron for reagents; D. J. Heard for critical reading of the manuscript and for discussions; I. Cheynel for expression of the TAFIIs and CPSF in the baculovirus system; Y. Lutz for antibodies; the Cell Culture group for HeLa cells; and R. Buchert and J.-M. Lafontaine for illustrations. J.-C.D. was supported by fellowships from the Ligue National Contre le Cancer and CNRS/Région Alsace. This work was supported by grants from CNRS, INSERM, Centre Hospitalier Universitaire Regional, Ministèe de la Recherche et Technologie, Fondation pour la Recherche Médicale and Association pour la Recherche contre le Cancer (L.T.) and from the NIH (J.L.M.).
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Dantonel, JC., Murthy, K., Manley, J. et al. Transcription factor TFIID recruits factor CPSF for formation of 3′ end of mRNA. Nature 389, 399–402 (1997). https://doi.org/10.1038/38763
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DOI: https://doi.org/10.1038/38763
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